D8276
DNA Polymerase I, Klenow Fragment from Escherichia coli
buffered aqueous glycerol solution
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About This Item
Recommended Products
grade
for molecular biology
form
buffered aqueous glycerol solution
mol wt
103 kDa
concentration
~3,000 units/mL
UniProt accession no.
foreign activity
Endonuclease, none detected
shipped in
wet ice
storage temp.
−20°C
Gene Information
Escherichia coli K12 ... polA(948356)
General description
DNA polymerase I yields two fragments (small and large) upon protease digestion. The large fragment (Klenow fragment) loses the 5′ exonuclease activity that is present in the intact holoenzyme. However, it retains both the polymerase 5′→3′ activity and the 3′→5′ exonuclease activity of the native enzyme.
Application
Suitable for:
- DNA sequencing by the Sanger dideoxy method
- Synthesis of the complementary strand of cDNA
- Filling in 5′-overhangs in double stranded DNA to form blunt ends
- Mutagenesis of DNA with second strand synthesis using oligonucleotides
- Labeling DNA by the random primer method
Components
DNA Polymerase I is supplied as a solution in 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 5 mM dithiothretol, and 50% glycerol (v/v) .
Unit Definition
One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min. at 37 °C.
Reconstitution
The enzyme solution may be diluted with 50 mM Tris-HCl, pH 7.5, 100 mM ammonium sulfate, 10 mM 2-mercaptoethanol, and 1 mg/ml bovine serum albumin.
Analysis Note
The activity is assayed in a reaction mixture containing 50 mM potassium phosphate (pH 7.5), 3 mM MgCl2, 1 mM 2-mercaptoethanol, 32.5 μM 32P-dATP, 32.5 μM dTTP, 62.5 μg/ml poly(dA-dT) and 0.01-1 unit enzyme.
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Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Resp. Sens. 1
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
Certificates of Analysis (COA)
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